| Background and ObjectiveAcute ischemic cerebral vascular disease is the most common type of cerebral infarction, and makes up60%-80%per cent of total cerebral vascular accident. It has a higher death rate and high disability rates. According to the relevant statistics about25%-75%recur in2-5year. Those make it with myocardial infarctions and malignant referred to as the three diseases with the highest death rate. Its clinical treatment is emphasis on early diagnosis, early treatment, early rehabilitation and early prevention of recurrence. All above Characteristics as well as high costs of treatment determine the importance of prevention, prophylactic therapy reduces the incidence and recurrence rates among high-risk populations.Butylphthalide,(dl-n-butylphthalide, dl-NBP), Can raise the level of cerebral vascular endothelial NO and PGI2, inhibit glutamate release, Can reduce the intracellular concentration of Ca2+, inhibition of free radicals and raise antioxidative enzyme activities and so on. Early treatment of acute ischemic stroke, gradually after the trial in chronic cerebral stroke and vascular dementia induced by, has good curative effect. In basic research have the exact molecular evidence, In view of its mechanism, intended to butylphthalide as prophylaxis, but similar studies currently rare.By far, S100B is the best way to reflect the degree of brain damage and prognosis of specific protein. Smac in mitochondria, and adjust the cysteine-aspartic acid protease (Caspase) path-dependent apoptosis. This research through the establishment of Wistar Rat model of acute focal cerebral ischemia of middle cerebral artery, observe butyl phthalide pretreatment on acute cerebral ischemia injury in cerebral protection effect and influence on the expression of Smac, S100B protein in brain tissue, explore its possible mechanisms.Materials and methods1、Animals and groups:33healthy male Wistar rats, weighing200-250g, Provided by the laboratory animal Center of Zhengzhou University; Randomly assigned to c Group (Group C), cerebral ischemia model group (Group M) and butyl phthalide pretreatment groups (Group NBP), each group of11.2、Cerebral ischemia model and pharmacological preconditioning:intragastric administration, Group NBP was treated by butyl phthalide liquid0.8g/kg of rat gavage, but the same volum of peanut oil for other two groups,Once a day for14days. And then, each one was treated by10%chloral hydrate (0.3ml/100g) intraperitoneal injection of anesthesia. for Group NBP and group M, Zea Longa improved intraluminal thread occlusion those Middle cerebral artery on the left side,but C not. After cerebral ischemia24hours, make them death.3、Detection of cerebral infarction volume:Randomly take brains from8rats in each group, Coronal slice5-6,then placed them in2%TTC in phosphate buffer solution, keep in dark place and a37℃water bath box lasts for30min. After dyeing, place all the brain slices in order, And photograph them both positive and negative. Calculation of cerebral infarction area and to the calculation of infarct size.4、Observation on pathological change in brain tissue:Select each group of8TTC staining typical infarct of brain tissue slice, dye them by HE staining, then observe on Penumbra area, finally take pictures.5、Expression of Smac, S100B protein in brain tissue: take brains from3rats in each group, then take100mg brain tissue, which blood was supplied by the left middle cerebral artery, to test their expression with Western Blotting. at last, chemiluminescence image and analysis the image, calculate the ratio grayscale.6、detect the moisture content of organization:by wet/dry weight ratio method. Brain tissue water content (%)=(wet-to-dry weight)/wet weight x100%.7、Statistical Methods:All experimental data were expressed by mean±standard error,(x±s), the groups were compared using single-factor analysis of variance, among groups were using the S-N-K method, the statistical software SPASS17.0for windows was used to analysis data, a=0.05.Result1、Comparison of pathological changes of brain tissue’Group C:Cell morphology and density normally, no edema. Group M:Brain tissue structure loosely, intercellular edema severely, arrange disorderly, varying degrees of neuronal degeneration and necrosis can be seen, the number of nerve cells significantly reduced, in the formation of cavities. Group NBP:Outcome measures more than minor changes, edema is not obvious, and fewer neuronal necrosis.2、 Comparison of cerebral infarction volume there is no brain infarcted area of rats in Group C; visible white brain infarcts can be seen in Group M and NBP, lines clearly.3、 Smac, S100B protein levels in brain tissue Relative to Group C, Smac, S100B protein level obviously increased inGroup M and Group NBP (P<0.05), while the two indexes in Group NBP were significantly lower than Group M (P<0.05).4、Comparison of water content in brain tissue Relative to Group C, water content obviously increased inGroup M and Group NBP (P<0.05), while it in Group NBP were significantly lower than Group M (P<0.05).Conclusion1、Suffering from acute hypoxic-ischemic damage, brain tissue necrosis occurs; Brain injuries and neuronal necrosis seriously, brain cell permeability change, apparent edema; the expression of Smac and S100B protein increase;2、Smac and S100B protein take part in brain ischemic injury of the pathological pathway;3、Butyl phthalide pretreatment can significantly reduce the ischemic brain cell damage, reduces infarct area and reduce brain edema in cerebral ischemic brain tissue;4、Butyl phthalide pretreatment significantly reduces apoptosis protein expression of Smac and S100B protein. |
PDF Full Text Request